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2017年09月21日 星期四
近期摘要
2015年第3期摘要
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闷黄过程中黄茶生化成分变化及其影响因子研究进展
滑金杰 1, 江用文 1, 袁海波 1*, 尹军峰 1*, 钟维标 2, 余书平 3,谢前途 4
1. 中国农业科学院茶叶研究所,浙江省茶叶加工工程重点实验室,国家茶产业工程技术研究中心,农业部茶树生物学与资源利用重点实验室,浙江 杭州 310008; 2. 平阳县天韵茶叶有限公司, 浙江 温州 325000; 3. 开化县林业特产局, 浙江 衢州 324000; 4. 平阳县特产站,浙江 温州 325400
摘要: 闷黄过程中, 黄茶中的生化成分在湿热作用下发生剧烈变化,如叶绿素因湿热引起的氧化降解反应而含量锐减; 儿茶素因氧化和异构化反应,其组分及比例发生明显改变; 氨基酸和可溶性糖含量亦有所变化等。 这些变化为形成黄茶黄汤黄叶、 醇爽口感等独特品质奠定了重要的物质基础。 文章对闷黄过程中,黄茶主要品质成分和微生物动态变化规律,以及闷黄工艺关键影响因子等进行了综述, 并对闷黄工艺研究趋势作了展望。
关键词: 黄茶; 闷黄;生化成分; 微生物; 影响因子
中图分类号: TS272.5            文献标识码: A           文章编号:1000-369X( 2015) 03-203-06
 
Review on the Changes of Biochemical Components and the Influencing Factors in Piling Process of Yellow Tea
HUA Jinjie1, JIANG Yongwen1, YUAN Haibo1*, YIN Junfeng1*, ZHONG Weibiao2, YU Shuping3, XIE Qiantu4
1. Tea Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Tea Processing Engineer of Zhejiang Province, National Engineering Technology Research Center for Tea Industry, Key Laboratory of Tea Biology and Resources Utilization Min istry of Agriculture, Hangzhou 310008, China; 2. Pingyang Tianyun Tea Co., Ltd., Wenzhou 325000, China; 3. Forestry specialty bureau of Kaihua county, Quzhou 324000, China; 4. Specialty station of Pingyang county, Wenzhou 325400, China
Abstract: During the piling process, dramatic changes of biochemical components in tea leaves are occurred under heat and humid effects: the content of chlorophyll drops sharply under oxidative degradation reactions, the composition and proportion of catechins are significantly changed under oxidation and isomerization reactions, the contents of amino acids and soluble sugar are also been changed. These changes provide an important material foundation for the unique quality formation of yellow tea: yellow soup, yellow leaves, mellow taste, etc. This paper reviewed the dynamic variation of quality biochemical compositions and microorganism in the piling process, the key influencing factors of piling process, as well as puts the prospects for the future research trends of piling technology.
Keywords: yellow tea, piling process, biochemical components, microorganism, influencing factors

 
安化千两茶香气品质特点分析
齐冬晴, 周跃斌,沈程文*, 蔡翔,王晓,余江意
教育部茶学重点实验室,湖南农业大学园艺园林学院,湖南 长沙 410128
摘要: 采用同时蒸馏萃取(SDE)技术,联合 GC-MS 鉴定方法分析了 9 个安化千两茶中的挥发性成分,结合感官审评初步探讨了这些成分的香气品质特点。感官审评结果表明,千两茶香气以松木香、陈香为主要特点,部分千两茶香气略带粽叶香或粗老气、烟气、陈气味,个别千两茶略带酸味。GC-MS分析结果表明,香气物质相对含量最多的为醇类(9个样品平均相对含量为21.97%);其次为酮类(20.11%)、醛类(18.69%)、杂氧化合物(18.39%)、脂肪酸类(12.71%)、烷烃类(1.70%)、含量最少的是杂环类(1.21%)。可见对千两茶香气起主要作用的是醇类、酮类、醛类和杂氧化合物。
关键词: 千两茶;香气;特点
中图分类号: TS272.5+4         文献标识码: A        文章编号:1000-369X(2015)03-209-08
 
Analysis of Aroma Quality Characteristic in An-hua Qian-liang Tea
QI Dongqing, ZHOU Yuebin, SHEN Chengwen*, CAI Xiang, WANG Xiao, YU Jiangyi
Tea Key Laboratory, Ministry of Education, College of Horticulture and Landscape, Hunan Agricultural University, Changsha 410128, China
Abstract: Simultaneous distillation extraction (SDE) technique combined with gas chromatography-mass spectrometry (GC-MS) identification method were applied to analyze the volatile components in nine kinds of An-hua Qian-liang tea, in which, the aroma characteristic of the components were discussed by sensory evaluation method. Results of sensory evaluation indicated that the main features of An-hua Qian-liang tea are fragrance of pine wood and stale flavour. Some An-hua Qian-liang teas possess a slight fragrance of reed leaves and aroma of coarse stems as well as the stale flavour and smoke. A few An-hua Qian-liang tea possesses a little sour taste. Results of GC-MS analysis indicated that alcohols, ketones, aldehydes, heterocyclic oxygen compounds play a major role on An-hua Qian-liang tea aroma. The alcohol content is the largest (21.97%), followed by aldehydes (20.11%), ketones (18.69%), heterocyclic oxygen compounds (18.39%), fatty acids (12.71%), lipids (1.70%) and heterocyclic (1.21%) compounds.
Keywords: Qian-liang tea, aroma, characteristic


冲泡过程中西湖龙井茶黄酮苷类浸出特性及滋味贡献分析
刘阳 1,2,陈根生 1,许勇泉 1,张英娜 1,2,尹军峰 1*
1. 中国农业科学院茶叶研究所 国家茶产业工程技术研究中心,浙江 杭州 310008; 2. 中国农业科学院研究生院,北京 100081
摘要: 本研究提出了一种基于 HPLC 的茶叶黄酮苷类物质检测方法,采用此方法分析了西湖龙井茶中黄酮苷类物质在不同冲泡条件下的浸出特性,并通过 Dot 值(浓度与阈值的比值)分析黄酮苷类物质对茶汤滋味的影响。结果表明:(1)该检测方法可以较好地分离和测定茶叶中 11 种黄酮苷类物质;(2)传统冲泡条件下,西湖龙井茶中以杨梅素-3-O-半乳糖苷(Myr-gala)和槲皮素-3-O-芸香糖苷(Que-rut)为主,随着冲泡温度的提高和时间的延长, 11 种黄酮苷类物质除了山柰酚-3-O-芸香糖苷(Kae-rut)外,都呈不同程度的增加,其中槲皮素-3-O-芸香糖苷(Que-rut)和槲皮素-3-O-半乳糖苷(Que-gala)的浸出速率较快,而杨梅素-3-O-鼠李糖苷(Myr-rha)和牡荆素-2"-O-鼠李糖苷(Vit-rha)的浸出速率较慢;(3)通过 Dot 值分析,槲皮素-3-O-芸香糖苷(Que-rut)、槲皮素-3-O-半乳糖苷(Que-gala)和杨梅素-3-O-半乳糖苷(Myr-gala)的 Dot 值均高于10,可能是茶汤滋味的重要贡献物质。
关键词: 冲泡;西湖龙井;黄酮苷类;浸出特性;滋味贡献
中图分类号:TS272.5+1; Q946.8    文献标识码:A    文章编号:000-369X (2015)03-217-08
 
Extracting Characteristics of Flavone and Flavonol Glycosides in Xihulongjing Tea under Different Brewing Conditions and Their Contribution to Tea Taste
LIU Yang1,2, CHEN Gensheng1, XU Yongquan1, ZHANG Yingna1,2, YIN Junfeng1*
1. Tea Research Institute, Chinese Academy of Agricultural Sciences, Engineering Research Cent er for Tea Processing, Hangzhou 310008, China; 2. Graduate school of Chinese academy of agricultural sciences, Beijing 100081, China
Abstract: The study proposed a standard analyzing method based on high performance liquid chromatography combined with ultraviolet (UV) to quantify extracting characteristics of flavone glycosides in Xihulongjing tea under different brewing conditions, and their contributions to taste was estimated by Dot factor (Dose-over-Threshold). Results indicated that: (1) This method can be used to separate and quantify eleven flavone and flavonol glycosides in tea; (2) In traditional brewing conditions, Myr-gala and Que-rut were the principal flavonol glycosides in Xihulongjing tea. With brewing temperature and time rising, eleven flavone and flavonol glycosides except Kae-rut increased variably with the raising of brewing temperature and the prolonging of brewing time, Que-rut and Que-gala leached fastly, while Myr-rha and Vit-rha leached slowly among 11 flavone and flavonol glycosides; (3)All of the Dot factors of Que-rut、 Que-gala and Myr-gala were higher than 10. According to the definition of Dot factor, they may be showed a significant contributor to tea taste.
Keywords: brew condition, Xihulongjing tea, flavone and flavonol glycosides, extracting characteristic, taste contribution


浙江省“十大名茶” 感官品质分析及风味轮的构建
张颖彬, 金寿珍, 王国庆, 于良子, 周苏娟, 刘新*, 鲁成银*
农业部茶叶质量监督检验测试中心,中国农业科学院茶叶研究所,农业部茶叶产品质量安全风险评估实验室(杭州), 浙江 杭州 310008
摘要: 研究收集了浙江省特别荣誉名茶西湖龙井和浙江省十大名茶 2013 年不同等级的 78 个样品,进行感官审评与理化分析。感官审评发现, 浙江省“十大名茶” 随着等级下降,茶叶风味品质逐渐降低,级差明显。 对不同样品的理化成分按照等级进行一般判别分析, 趋势较好。文章对浙江省“十大名茶”的香气、滋味类型,香气和滋味中的缺陷类型与数量进行了归纳与统计。在此基础上,总结出浙江省“十大名茶”品质特征,对各个名茶的香气、滋味类型进行了解析与重构,首次绘制完成了浙江省“十大名茶” 香气、滋味风味轮。
关键词: 浙江省“十大名茶” ; 感官评价; 感官风味; 风味轮
中图分类号: TS272.5+1         文献标识码: A          文章编号: 1000-369X( 2015)03-225-08
 
Flavor Analysis and Flavor Wheel Establishment of Ten Top Famous Tea in Zhejiang Province
ZHANG Yingbin, JIN Shouzhen, WANG Guoqing, YU Liangzi, ZHOU Sujuan, LIU Xin*, LU Chengyin*
Tea Quality and Supervision Testing Center, Ministry of Agriculture,Tea Research Institute, Chinese Academy of Agricultural Sciences, China National Laboratory for Tea Safety Risk Assessment, Hangzhou 310008, China
Abstract: 78 samples of different grades (including top famous tea and special famous tea in Zhejiang province)were collected in this research. Through the sensory evaluation, an obvious flavor changes between different grades were found. While processing the data of chemical compounds in different grade samples with discriminant analysis, a well judgment can be shown, which is essentially in agreement with the sensory evaluation result. The quantity of samples with different aroma and taste types was counted, and then the characteristics of each tea were summarized.
Keywords: top famous tea, sensory evaluation, flavor, flavor wheel

 
TF1 与 EGCG 在 Caco-2 细胞中的吸收规律研究
孟庆,屠幼英*
浙江大学农业与生物技术学院茶学系,浙江 杭州, 310058
摘要: 为了探究茶黄素单体( TF1)与表没食子儿茶素没食子酸酯( EGCG) 在生物体中的吸收规律,本实验建立了体外 Caco-2 单层细胞模型, 模拟小肠对 TF1 与 EGCG 的吸收,并研究了浓度和时间对吸收的影响。 结果表明, 在 10~100 µmol·L-1 范围内, TF1 与 EGCG 在 Caco-2 单层细胞模型中的吸收呈现表观渗透系数 Papp随浓度增加而上升的规律。但两者的外排率也呈现同样的规律,并且外排率上升的幅度均大于二者吸收率上升的幅度。由于 TF1 与 EGCG 在细胞单层模型中的 Papp 均小于 1×10-6 cm·s -1,说明两者都属于难吸收的药物,但是 TF1 在 Caco-2 细胞模型中的吸收率高于 EGCG。因其外排比均大于 2,说明两者在细胞模型上的外排是被动转运。从吸收时间看, TF1 的外排规律与 EGCG 一致。
关键词: TF1; EGCG; Caco-2 细胞模型;吸收规律
中图分类号:TS272;Q946.84+1    文献标识码:A     文章编号:1000-369X(2015)03-233-06
 
Study on Absorption of Theaflavin and EGCG in Caco-2 Cell Model
MENG Qing, TU Youying*
Department of tea science, Zhejiang University, Hangzhou 310058, China
Abstract: In order to explore the absorption regularity of TF1 and EGCG in the organism. This research used the Caco-2 cell monolayer model in vitro to simulate the absorption of TF1 and EGCG in the small intestine. The influences of concentration and time on the absorption regularity of TF1 and EGCG in Caco-2 cell monolayer model were investigated. The results illustrated that the absorption of TF1 and EGCG in Caco-2 model showed that the apparent permeability coefficient raised with the increasing of concentrations of two compound in the range of 10~100 µmol·L-1. The efflux rates of the TF1 and EGCG showed the same rules as absorption. However, the increasing range of efflux rate was higher than that of absorption rate. The values of Papp about TF 1 and EGCG in the cell model were lower than 1×10-6 cm·s -1, which indicated that both of them belonged to the kind of drugs which were difficult to absorb. However, with comparing the absorption rate, TF1 was higher than EGCG in this model. Both of efflux transport showed the passive process because the efflux rates of TF 1 and EGCG were larger than 2 and higher than in cell model. Because the efflux regulation of TF 1 was in accordance with EGCG by temporal variation, it suggested that both of them were the substrate of the same efflux protein.
Keywords: TF1, EGCG, Caco-2 model, absorption regularity
 
 
茶多酚防治 2 型糖尿病的分子机理研究进展
高媛圆 1, 毛立民 2, 徐平 1,王岳飞 1*
1.浙江大学茶学系,浙江 杭州 310058; 2. 浙江省茶叶学会,浙江 杭州 310029
摘要: 茶多酚作为天然功能性物质能有效防治 2 型糖尿病,但是其具体作用机制仍不太清楚。研究表明它的作用机理主要是通过调控糖代谢平衡: 包括降低 α-糖苷酶、 α-淀粉酶等双糖酶活性, 调控胰岛素信号传导途径及 AMPK 途径中多个分子靶点的磷酸化水平,影响酶与转录因子的活性与表达,减弱胰岛素抵抗及糖异生作用,促进胰岛素的合成与分泌,促进靶组织对葡萄糖的吸收与利用等;同时,茶多酚能够调控脂代谢,通过抑制脂质合成与积累的相关酶的活性及转录因子的表达,促进脂质的氧化代谢,减轻脂代谢紊乱;另外,茶多酚的抗氧化与抗炎作用可以有效减轻组织细胞的氧化伤害和炎症损伤,减少细胞凋亡。
关键词: 茶多酚; EGCG; 2 型糖尿病;分子机理
中图分类号: Q946.84+1;R587.1  文献标识码:A  文章编号:1000-369X(2015)03-239-09
 
Advances in Molecular Mechanisms of Tea Polyphenols in Preventing Type 2 Diabetes Mellitus
GAO Yuanyuan1, MAO Limin2, XU Ping1, WANG Yuefei1*
1. Department of Tea Science, Zhejiang Univerisity, Hangzhou 310058, China; 2. Zhejiang Tea Science Society, Hangzhou 310029, China
Abstract: As natural functional materials, tea polyphenols have been shown to effectively prevent the development of type 2 diabetes mellitus (T2DM), but the mechanism of action is still unclear. For further study, this article summarizes the molecular mechanisms implicated in the beneficial metabolic effects of tea polyphenols. In the respect of glycometabolism, tea polyphenols significantly reduce the absorption of simple sugars by inhibiting disaccharidases (α-amylase and α-glucosidase). Then, tea polyphenols can ameliorate insulin resistance and inhibit gluconeogenesis via insulin signaling and AMPK pathway, including regulation of the phosphorylation and expression of protein kinases, and relevant transcription factors. In addition, tea polyphenols stimulate glucose uptake and utilization by increasing insulin secretion. Meanwhile, tea polyphenols showed great effects on improving lipid metabolic disorder by suppressing lipid synthesis and accumulating the activity of related enzyme and the expression of transcription factors as well as stimulating the oxidative metabolism of fat and lipid and suppressing the lipidosis. Moreover, tea polyphenols showed antioxidative and anti-inflammatory capability to ameliorate cell damage and apoptosis induced by oxidation and inflammation.
Keywords: tea polyphenols, EGCG, type 2 diabetes mellitus, molecular mechanism

 
施肥结构对茶园土壤氮素营养及茶叶产量品质的影响
朱旭君 1,王玉花 1,张瑜 1,肖润林 2,黎星辉 1*
1.南京农业大学茶叶科学研究所, 江苏 南京 210095; 2. 中国科学院亚热带农业生态研究所, 湖南 长沙 410125
摘要:通过设置施氮水平、有机肥与化肥配施比双因素田间试验,研究了不同施肥结构对茶园土壤有机质和氮素营养、 春茶产量和品质的影响。 结果表明,与不施肥处理相比,各种施肥处理均能有效提高茶园土壤有机质和全氮含量,且均在 900 kg·hm-2 的施氮水平下 75%有机肥+25%化肥处理效果最为显著; 施肥可以显著提高春茶芽叶密度、百芽重等产量指标; 有机肥比例高于 25%的氮素处理能显著提高春茶的品质指标,有机肥比例 50%以上时效果更好。
关键词: 施肥结构; 茶园; 氮素营养; 茶叶产量; 茶叶品质
中图分类号: S571.1;S157.4+1     文献标识码:A     文章编号:1000-369X(2015)03-248-07
 
Effects of Different Fertilizer Application Systems on Nitrogen Nutrition in Tea Garden Soil and Yield-Quality of Tea Plant
ZHU Xujun1, WANG Yuhua1, ZHANG Yu1, XIAO Runlin2, LI Xinghui1*
1.Institute of Tea Science, Nanjing Agriculture University, Nanjing 210095, China; 2. Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China
Abstract: Through field test by setting factors of nitrogen level and organic/chemical fertilizer ratio, we studied the effects of different fertilizer application systems on organic matter and nitrogen nutrition in tea garden soil, as well as tea yield & quality were investigated. The results showed that contents of organic matter and total nitrogen were significantly increased under all various fertilizer treatments, and applying 75% organic manure combined with 25% chemical fertilizers with 900 kg·hm-2 high nitrogen level exhibited the most remarkable increasing effects. Compared with no fertilization treatment, indicators of tea production including the bud-leaf density, 100-bud weight were increased under all various fertilizer treatments. Furthermore, the quality indicators of spring tea were dramatically increased under such fertilizer treatments that organic manure more than 25%, the tea quality improved more while organic manure was more than 50%.
Keywords: fertilizer application systems, tea garden, nitrogen nutrition, tea production, tea quality


 茶树修剪物对茶园土壤真菌群落多样性影响研究
袁赛艳, 陈暄, 房婉萍, 王玉花,朱旭君, 黎星辉*
南京农业大学茶叶科学研究所,江苏 南京 210095
摘要: 茶树修剪物直接还田是茶园管理中常见的栽培措施。 本文通过提取不同处理的土壤样品总 DNA, 以未处理土壤样品为对照, 利用变性梯度凝胶电泳(PCR-DGGE), 研究添加茶树修剪物对茶园土壤真菌群落结构多样性的影响。结果显示,各处理土壤样品的 DGGE 图谱中发现许多公共条带,这些共有条带主要属于Pezizomycotina 门、 Basidiomycota 门、 Mortierellomycotina 门和 Mucoromycotina 门; 通过 DGGE 的条带数和亮度进行真菌群落多样性各项指标的比较, 显示各处理的丰富度指数和香农-威纳指数均低于对照,表明茶树修剪物的降解产物和茶树本身固有的酚类物质对茶园的真菌群落有一定的抑制作用。
关键词: 茶树修剪物; PCR-DGGE; 群落结构; 多样性
中图分类号:S571.1;S154.3   文献标识码:A   文章编号:1000-369X(2015)03-255-08
 
Effect of Tea Plant Pruning Material on Fungi Community Diversity in Tea Plantation Soil
YUAN Saiyan, CHEN Xuan, FANG Wanping, WANG Yuhua, ZHU Xujun, LI Xinghui*
Tea Research Institute, Nanjing Agricultural University, Nanjing 210095, China
Abstract: Returning of pruning material to the field is a common agronomic practice performed in tea plantation. By extracting the total DNA of soil in various treatments, the effects of tea pruning material on fungal community structure and diversity in tea garden soils were investigated by using PCR amplification and denaturing gradient gel electrophoresis (DGGE), the original soil without any supplement was set as control. Many common bands were detected from each soil samples by DGGE, and those bands that showed highest sequence homology to Pezizomycotina, Basidiomycota, Mortierellomycotina and Mucoromycotina. The number and brightness of bands from DGGE presented the community diversity of fungi under different treatments. The results showed the richness and Shannon index were lower than the control, maybe the degradation products and phenolic compounds of pruning material have some inhibitory effect on fungi community diversity.
Keywords: tea plant pruning material, PCR-DGGE, community structure, diversity
 
 
 不同类型茶园土壤腐殖质剖面分布特征研究
王峰 1,陈玉真 1,尤志明 1*,吴志丹 1,江福英 1, 翁伯琦 2, 张文锦 1
1福建省农业科学院茶叶研究所,福建 福安 355015; 2. 福建省农业科学院农业生态研究所,福建 福州 350013
摘要:采用野外调查和室内分析相结合的方法,对武夷山市 5 种主要土壤类型 (黄壤、红壤、潮砂土、高山草甸土、紫色土)茶园的土壤腐殖质剖面分布特征进行研究。 结果表明, 茶园土壤腐殖质各组分含量基本随着土层加深呈减少趋势。不同类型茶园土壤腐殖质的含量和组成存在显著差异,高山草甸土茶园土壤腐殖质及各组分的含量均显著高于其他土壤类型,黄壤和紫色土次之,红壤最低。土壤腐殖质组成中,均以胡敏酸比例最高(57.06%~79.76%),除高山草甸土外,各土壤中富里酸比例均大于胡敏酸。 茶园土壤富里酸的色调系数(△logK) 和光密度值(E4/E6)均大于胡敏酸,高山草甸土富里酸和胡敏酸的△logK 和 E4/E6 值最低,土壤腐殖化复杂程度最高;红壤的富里酸和胡敏酸的△logK 和 E4/E6 值最高,土壤腐殖质复杂程度及化学稳定性低。相关分析表明,土壤腐殖质组成与土壤有机碳、酚类含量、全氮、土壤容重和孔隙度存在显著相关性,与土壤 C/N、pH 和含水率相关性不明显。可见,茶园土壤腐殖质均以胡敏素为主,高山草甸土为胡敏酸型,其余土壤为富里酸型。
关键词: 茶园; 土壤类型;土壤腐殖质;剖面分布
中图分类号: S571.1; S153.6+22   文献标识码:A   文章编号:1000-369X(2015)03-263-08
 
The Vertical Characteristics of Soil Humus in Different Soil Types of Tea Garden
WANG Feng1, CHEN Yuzhen1, YOU Zhiming1*, WU Zhidan1, JIANG Fuying1, WENG Boqi2, ZHANG Wenjin1
1. Tea Research Institute, Fujian Academy of Agricultural Sciences, Fu’an 355015, China; 2. Agriculture Ecology Institute, Fujian Academy of Agricultural Sciences, Fuzhou 3 50013, China
Abstract: Based on the field survey and laboratory analysis, the soil humus composition in five soil types (yellow soil, red earth, moisture sandy soil, alpine meadow soil and purple soil) of tea garden in Wuyishan city were investigated. The results showed that the content of humic acid(HA), fulvic acid(FA) and Humin(HM) decreased with the soil depth. There were significant differences of humus composition among different soil types. The humus composition and carbon level in alpine meadow soil of tea garden were significantly higher than those in other soil types, followed by those in yellow soil and purple soil, the lowest was in red soil. Humin was the main composition of humus (57.06%-79.76%). Except alpine meadow soil, the proportion FA were more than those in HA. The △log K and E4/E6 value of FA was higher than HA. The △log K and E4/E6 value in alpine meadow soil and red soil were the lowest and highest, respectively. The soil humus composition were significantly correlated with soil organic carbon and phenol contents, total N, soil bulk density and soil porosity, but no significant correlated with C/N, pH value and soil moister. Thus, the major type of humus was humin in all five kinds of tea garden soils, alpine meadow soil belonged to the humic acid type soil, and others belonged to fulvic acid enriched soil.
Keywords: tea garden, soil types, soil humus, profile distribution

产多酚氧化酶茶树内生真菌的筛选及产酶条件优化
张婉蓉 1,巫婷玉 2,杨民和 1,2*
福建师范大学生命科学学院, 福建 福州 350108; 2. 工业微生物教育部工程研究中心 福建师范大学, 福建 福州 350108
摘要:以愈创木酚、α-萘酚、没食子酸、L-酪氨酸和单宁酸 5种酚类物质为底物,采用鉴别培养基筛选法从14 株茶树内生真菌中初筛获得4株产多酚氧化酶的真菌。根据变色圈的大小、颜色深浅和摇瓶发酵的结果复筛获得产多酚氧化酶能力较强的菌株 CSN-13。对菌株 CSN-13 产酶营养条件进行初步分析,结果表明,在供试的 6 种碳源物质中,以麸皮对菌株 CSN-13 产多酚氧化酶的促进作用最为明显;供试的 5 种氮源物质中,以硝酸铵的促进作用最为明显;在发酵培养基中添加茶水,对产酶有明显的促进作用。采用正交设计对CSN-13 产酶发酵培养基进行初步优化,优化后的培养基配方为:麸皮(40 g·L-1)、硝酸铵(15 g·L-1)、茶水(4 g·L-1)、KH2PO4(2 g·L-1)、MgSO4·7H2O(0.5 g·L-1)、无水CaCl2(0.075 g·L-1)、CuSO4·5H2O(0.01 g·L-1)。采用优化后的培养基,菌株CSN-13在28℃下培养5 d,酶活力达到241 U·mL-1·min-1,比优化前提高 8.5 倍。茶树内生真菌菌株 CSN-13 及其发酵产酶培养基的研究为多酚氧化酶的进一步开发打下了基础。
关键词: 茶树;内生真菌; 多酚氧化酶; 菌株筛选; 培养基优化
中图分类号:S571.1; Q949.32   文献标识码:A    文章编号:1000-369X(2015)03-271-10
 
Screening and Culture Medium Optimization of Polyphenol Oxidase Producing-Fungi from Endophytes of Tea Plant (Camellia sinensis)
ZHANG Wanrong1, WU Tingyu2, YANG Minhe1,2*
1.College of Life Science, Fujian Normal University, Fuzhou 350108, China; 2. Engineering Research Center of Industrial Microbiology Affiliated to Ministry of Education, Fujian Normal University, Fuzhou 350108, China
Abstract: Five kinds of phenolic materials, including guaiacol, alpha-naphthol, gallic acid, L-tyrosine and tannic acid, were used as the substrates to screen polyphenol oxidase-producing fungal strains. Four out of 14 endophytic fungal strains of tea plant (Camellia sinensis) were selected by agar plate screening methods. Strain CSN-13 was obtained for its color ring size and color depth, which owned a powerful production capacity of polyphenol oxidase. A preliminary analysis for nutritional components of strain CNS-13 was conducted. The results showed that among 6 kinds of carbon sources, wheat bran was the best for its promoting effects upon polyphenol oxidase production. Ammonium nitrate showed the most significant promotion among 5 kinds of nitrogen sources. Enzyme production was significantly promoted when tea infusion was added into the fermentation medium. With the adoption of orthogonal design, the fermentation medium for enzyme production was preliminarily optimized. The optimized medium contains wheat bran (40 g·L-1), ammonium nitrate (15 g·L-1), tea (4 g·L-1), KH2PO4 (2 g·L-1), MgSO4·7H2O (0.5 g·L-1), CaCl2 (0.075 g·L-1), and CuSO4 could reach 241 U·mL-1·min-1 under culture conditions with 28℃ temperature for 5 days, which was 8.5 times higher than that of before optimization. Strain CSN-13 and the fermentation medium in present study offered an alternative for the further development of polyphenol oxidase.
Keywords: Camellia sinensis, endophyte, polyphenol oxidase, fungal strain screening, medium optimization
 
 
不同 pH 条件下 TSs 的形成机理及其与TFs 的竞争性形成研究
徐斌 1,2, 江和源 1*,张建勇 1, 杨刘艳 1,2, 刘千录 1,2
1.中国农业科学院茶叶研究所 农业部茶树生物学与资源利用重点实验室 浙江省茶叶加工工程重点实验室,浙江 杭州 310008; 2. 中国农业科学院研究生院,北京 100081
摘要:采用液态发酵的方式,结合不同的 pH 条件,通过酶促氧化儿茶素生成相应聚酯型儿茶素( Theasinensins, TSs)或茶黄素( Theaflavins, TFs)的单体物质来深入探讨 TSs 的形成机理及其与 TFs 的竞争性形成。结果表明, 酸性条件下, TSs 的单体物质能够持续生成,而且一直积累;中性和碱性条件下生成的 TSs 单体物质达到最大值的时间很短, 稳定性较差。不同 pH 条件下,由于自身氧化还原电位值的高低,各个儿茶素单体之间被氧化而消耗的速率有所差别, 而且基于生成 TSs 和 TFs 需要共同的儿茶素底物,因此两者的形成具有竞争性。pH=6 和中性条件下 EGCG 和 EGC 转化形成 TSs 单体的量高于形成 TFs 的量,而 pH=5 水平下则相反。
关键词: 儿茶素;聚酯型儿茶素;茶黄素;形成机理;竞争性
中图分类号: TS272; Q946.84+1 文献标识码: A   文章编号:1000-369X(2015)03-281-09
 
Formation Mechanism of TSs and Competitive Formation Between TSs and TFs under Various pH
XU Bin1,2, JIANG Heyuan1*, ZHANG Jianyong1, YANG Liuyan1,2, LIU Qianlu1,2
1Tea Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Tea Plants Biology and Resources Utilization of Agriculture Ministry, Key Laboratory of Tea Processing Engineering of Zhejiang Province, Hangzhou 310008, China; 2. Graduate School of Chinese Academy of Agricultural Sciences, Beijing 100081, China
Abstract: To illustrate the formation mechanism of theasinensins (TSs) and the competitive formation between TSs and theaflavins (TFs), catechins(EGCG, EGC, EC and ECG) were oxidized to form TSs monomers or TFs monomers by enzymatic oxidation and the experiments were performed by liquid fermentation under different pH values. The results showed that the TSs monomers were synthesized and accumulated continuously under acidic conditions, while the peaks of TSs monomers appeared in a short time but showed poor stability under neutral or alkaline conditions. Due to the magnitudes of the oxidation reduction potential (ORP) are variable for catechins, the enzymatic oxidative rates of catechins are different under different pH conditions. For the reason that there was a competition between the formation of TSs and TFs: the content of TSs monomers transformed by EGCG and EGC was higher than TFs monomers under the neutral condition or pH=6, however, conversely under the condition pH=5.
Keywords: catechin, theasinensins, theaflavins, formation mechanism, competitiveness
 

茶树花粉中丙酮酸脱氢酶( CsE1α)的定位分析及其启动子的克隆与表达
杜昱林, 王伟东, 王玉花*,黎星辉
南京农业大学茶叶科学研究所, 江苏 南京 210095
摘要: 根据以往试验获得的 CsE1α 的 cDNA 全长序列,利用 TAIL-PCR 克隆 CsE1α 启动子。测序验证与生物信息学分析后发现,该启动子片段长 336 bp,含有 2 个 CAAT-box,2 个 TATA-box,2 个 GATA-box,1个 LTR,1 个 G-box 等顺式作用元件。构建载体转入洋葱内表皮细胞瞬时表达,启动子可启动下游报告基因,使荧光蛋白表达于整个细胞,表明所克隆的启动子具有启动功能。 CsE1α 与 GFP 融合蛋白瞬时表达表明 CsE1α 定位于线粒体。本实验为下一步转基因拟南芥稳定表达,进一步研究CsE1α基因的表达调控,探讨茶树花粉抗寒的分子生物学机理奠定基础。
关键词: 茶树; 丙酮酸脱氢酶; 启动子; 亚细胞定位; 抗寒
中图分类号:S571.1; Q946.5     文献标识码: A      文章编号: 1000-369X(2015)03-290-09
 
Subcellular Localization of CsE1α as well as Cloning and Expression of Its Promoter from the Pollen of Camellia sinensis
DU Yulin, WANG Weidong, WANG Yuhua*, LI Xinghui
Tea Research Institute, Nanjing Agricultural University, Nanjing 210095, China
Abstract: In this paper, full-length cDNA is identified for designing the gene-special primers in TAIL-PCR to clone the promoter of CsE1α. By sequencing and bioinformatic analyzing, we observed two CAAT-box, two TATA-box, two GATA-box, one LTR and one G-box located in the 336 bp promoter region. After constructing and transferring the transient expression vectors into the onion epidermal cells, subcellular localization of CsE1α-GFP fusion proteins is verified in mitochondria, while the promoter can activate downstream gene expressing in entire cell. This experiment may provide useful information for the subsequent stable expression in transgenic Arabidopsis and the further investigation on the expression and regulation of CsE1α gene as well as the investigation on the molecular mechanism of cold resistance in pollen of Camellia sinensis.
Keywords: Camellia sinensis, pyruvate dehydrogenase, promoter, subcellular localization, cold resistance
 
 
茶文化资源类型及业态范式研究
沈学政, 苏祝成, 王旭烽
浙江农林大学艺术设计 人文·茶文化学院,浙江 临安 311300
摘要:中华茶文化通过与农业融合反哺地方经济,形成茶文化产业。 本文采用文化资源基础理论,将茶文化资源分为五大主要类型,并进一步细分为 15 种基本类别和 13 种茶文化产业类型。随后以福建省、武夷地区、大益品牌为案例,分别从一个省、一个地区、一个企业的角度总结茶文化资源 3 种产业开发范式,即比较优势模式、点轴开发模式和产业延伸模式,探讨茶文化资源综合开发对于区域经济和茶业转型升级的意义。
关键词: 茶文化;文化资源; 资源类型; 业态范式
中图分类号:S571.1;G124        文献标识码: A         文章编号:1000-369X(2015)03-299-08
 
Research on the Type and Mode of the Development of Tea Culture Resource
SHEN Xuezheng, SU Zhucheng, WANG Xufeng
Tea Culture Institute, Zhejiang A&F University, Lin′an 311300, China
Abstract: Chinese tea culture converged the local economy and agriculture to form the tea cultural industry. Based on the theory of cultural resource, this thesis divided the tea culture heritage resources into 5 major types. And further more, we subdivided it into 15 basic categories. Meanwhile, in view of the different types of tea cultural heritage resources, we put forward 13 main development mode, which promote the regional economy and advance the tea industry transformation and upgrading. Then, taking Fujian Province, Wuyi area, TAETEA as the case, respectively, we summed up 3 kinds of development mode of tea cultural heritage resource, from a province, a region, a business point of view, namely comparative advantage model, point axis development model and the extended model industry.
Keywords: tea culture, heritage resource, resource type, development mode
 
 

 

 

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